The current era of multi-drug resistance is having dramatic impacts on human health, public health and the practice of clinical medicine. Improved technology is needed to provide prompt and accurate information to treating clinicians for best patient management. The overarching goal on this project is to develop, validate and translate a rapid detection and expression array, for the clinical laboratory setting, to comprehensively characterize antibiotic resistance in important pathogens (Pseudomonas aeruginosa, Klebsiella pneumonia, Acinetobacter baumannii, Staphylococcus aureus, and Streptococcus pneumoniae) in clinical samples. Such a system would assist decision-making with respect to antibiotic use, help slow the onset of additional resistance, and improve disease surveillance. This array will provide pathogen detection, markers for presence and expression of antibiotic resistance genes/mutations and, where applicable and pertinent, strain-typing information that can also be used for diagnostic and in-hospital surveillance. The logical flow of the research plan will include capturing all current knowledge on antibiotic resistance characteristics for target pathogens; conducting additional analyses to further knowledge on species signatures and antibiotic resistance characteristics; developing novel PCR assays for these targets; validating developed assays, both individually and together on the high-density format array (using large strain and clinical sample repositories); developing the bioinformatic tools necessary to easily read and interpret array findings; and finally transferring developed technology to a clinical laboratory for evaluation. The project team has expertise from translational research, genomic analysis, clinical-based research, molecular epidemiology, public health and medical biotechnology development. Previously established collaborations among the team members will allow for immediate implementation of the proposed activities and early impacts on this grave threat. Clinical medicine and public health are facing the growing crisis of multi-drug resistance in common pathogens. More informative and rapid diagnostics are urgently needed to provide clinicians and public health officials the information necessary to appropriately respond to these infections. This project will develop, validate and translate a rapid high-density antibiotic-resistance detection system for five high-priority multi-drug resistant pathogens; such a system would assist decision-making with respect to antibiotic use, help slow the onset of additional resistance, and improve disease surveillance.